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Precision of genotyping of Epstein-Barr virus by polymerase chain reaction
using three gene loci (EBNA-2, EBNA-3C, and EBER): predominance of type A
virus associated with Hodgkin's disease [published erratum appears in Blood
1993 Oct 1;82(7):2268]
JC Lin, SC Lin, BK De, WC Chan, BL Evatt and WC] Chan WP [corrected to Chan
Molecular Biology Section, Centers for Disease Control, Atlanta, GA 30333.
To precisely determine the genotype of Epstein-Barr virus (EBV) in
Hodgkin's disease (HD), we simultaneously analyzed three divergent gene
loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The
primers designed to amplify these three gene loci encompass either
type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific
point mutations (EBER) that identify the virus strain based on the sizes of
the polymerase chain reaction (PCR)-amplified products or the mobility
shifts in single-strand conformation polymorphism analysis. The locations
of point mutations were identified by direct sequencing of the
PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good
correlation between EBNA-2 and EBNA-3C typing results. In contrast,
approximately 33% of the cell lines analyzed maintained type A sequences in
EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER
region. Data obtained from analysis of cell lines served as a reference for
studying HD samples. EBV DNA was detected in about 70% of HD. Among the
EBV-positive samples, 56% were associated with type A virus, 13% with type
B, and 31% with dual viral sequences. Thus, type A virus is predominant in
HD. Based on the histology, the frequencies of EBV positivity were 83%,
71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte
predominance, respectively. The detection of high frequency of both type A
and B sequences in HD may provide a lead in investigating the role of dual
viral infection in EBV pathogenesis.
Volume 81,
Issue 12,
pp. 3372-3381,
06/15/1993
Copyright © 1993 by The American Society of Hematology
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